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Applications of nucleic acids have developed recently to provide solutions for biosensors, diagnostic tools and as platforms for the assembly of complex structures. These developments have been possible as their base sequence can be used to assemble precise structures following simple and predictable rules. Self-assembled DNA can then be amplified using polymerase chain reaction (PCR) and this ultimately enables the preparation of synthetic nucleic acids. Their use as molecular tools or DNA-conjugates has recently been enhanced by the addition of other groups including enzymes, fluorophores and small molecules. Written by leaders in the field, this volume describes the preparation and application of these DNA-conjugates. Several have been used as sensors (aptamers, riboswitches and nanostructures) based on the ability of nucleic acids to adopt specific structures in the presence of ligands, whilst others link reporter groups such as proteins or fluorophores to RNA or DNA for detection, single molecule studies, and increasing the sensitivity of PCR. The book is relevant to researchers in areas related to analytical chemistry, chemical biology, medicinal chemistry, molecular pharmacology, and structural and molecular biology.
This book focuses on the basic electrochemical applications of DNA in various areas, from basic principles to the most recent discoveries. The book comprises theoretical and experimental analysis of various properties of nucleic acids, research methods, and some promising applications. The topics discussed in the book include electrochemical detection of DNA hybridization based on latex/gold nanoparticle and nanotubes; nanomaterial-based electrochemical DNA detection; electrochemical detection of microorganism-based DNA biosensors; gold nanoparticle-based electrochemical DNA biosensors; electrochemical detection of the aptamer-target interaction; nanoparticle-induced catalysis for DNA biosensing; basic terms regarding electrochemical DNA (nucleic acids) biosensors; screen-printed electrodes for electrochemical DNA detection; application of field-effect transistors to label free electrical DNA biosensor arrays; and electrochemical detection of nucleic acids using branched DNA amplifiers.
The aim of this research was to develop stable dye-labelled oligonucleotidenanoparticle conjugates for the detection of DNA related to disease using surface enhanced resonance Raman scattering (SERRS) spectroscopy. In order to achieve this, both gold and silver nanoparticle conjugates were successfully functionalised with thiol- and thioctic acid-modified oligonucleotides. To investigate their viability as biosensors, the stability of the conjugates and their hybridisation efficacy was investigated. It was discovered that the thioctic acidmodified oligonucleotide-nanoparticle conjugates remain stable under these conditions for a substantial period of time compared to thiol-modified oligonucleotide nanoparticle conjugates. The nanoparticle conjugates were functionalised for use in surface enhanced resonance Raman scattering (SERRS) spectroscopy by the incorporation of an isothiocyanate dye molecule. Not only were the conjugates SERRS active, they were also proven to exhibit enhanced stability for both thiol- and thioctic acid-modified oligonucleotide nanoparticle conjugates. The thioctic acid-modified oligonucleotide nanoparticle conjugates were investigated for their use as sensors to detect specific DNA sequences. Both gold and silver nanoparticle conjugates were shown to retain their biological activity by hybridising with complementary DNA sequences. This was monitored colorimetrically, due to the characteristic colour changes associated with aggregation of gold and silver nanoparticles, and by monitoring the hybridisation-induced red-shift of the surface plasmon band by UV-Vis spectroscopy. The hybridisation-induced aggregation methodology was also extended to dye-labelled nanoparticle conjugates that were investigated by SERRS. A method for characterising oligonucleotide-nanoparticle conjugates using a fluorescent DNA intercalator, SYBR green I, was developed. This study showed that SYBR green I can be used as a rapid method for determining the melting profiles of individual nanoparticle conjugates and the hybridisation efficiency of the immobilized oligonucleotides.
Biological DNA Sensor defines the meaning of DNA sensing pathways and demonstrates the importance of the innate immune responses induced by double stranded DNA (dsDNA) through its influencing functions in disease pathology and immune activity of adjuvants for vaccines. Though discussed in specific subsections of existing books, dsDNA and its immunogenic properties has never received the complete treatment given in this book. Biological DNA Sensor approaches the impact of dsDNA's immunogenicity on disease and vaccinology holistically. It paints a complete and concise picture on the topic so you can understand this area of study and make more informed choices for your respective research needs. Chapters are authored by researchers who are renowned for their research focus, ensuring that this book provides the most complete views on the topics. Multi-authored by a distinguished panel of world-class experts Ideal source of information for those wanting to learn about DNA sensing Provides in-depth explanations of DNA sensing pathways and the innate immune system, bridging the gap between them
DNA Sensors and Inflammasomes, Volume 625, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. New sections in this release include Phosphorylation and dimerization of STING and IRF3, cGAS enzymology, Synthesis and identification of immuno-stimulatory CDNs, Tracking cGAS activity/ cGAMP formation using SPR/NMR, Using an enzyme coupled assay to track cGAS activity under steady states, Tracking the polymerization of DNA sensors, inflammasome receptors, and downstream signaling partners using FRET, NLRC4 structure, Tracking TREX1 activity, DNA association and dissociation kinetics of PARP1, and more. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology series Includes the latest information on DNA sensors and inflammasomes
The development of highly sensitive and selective DNA sensors has fuelled applications in a wide range of fields including medical diagnostics, forensics, biodefense, food contamination and environment monitoring. Recently, conjugated polymers (CPs) have drawn attention as attractive novel materials for biosensors due to their tuneable optical and electronic properties and ability to respond to minor changes in their environment. The aim of this research was to develop miniaturised optical DNA sensors based on photoluminescent CPs. Investigations towards the effect of systematically substituting carboxylic acid functionalized CPs with polar methoxy, mono-ethylene glycol, di-ethylene glycol and tri-ethylene glycol sidechains on the solubility, optical and electrochemical properties were conducted. The resultant CPs were characterised using a range of techniques such as nuclear magnetic resonance (NMR), fourier infrared spectroscopy (FTIR), ultra-violet (UV-Vis) visible spectroscopy and fluorescence spectroscopy. Furthermore, three different novel optical DNA sensors on magnetic beads were designed and developed using the synthesized CPs. DNA hybridisation was monitored using fluorescence resonance energy transfer (FRET) and superquenching readout methodologies. These sensors exhibit desirable characteristics such as ease of handling (through magnetic deployment). All the three different sensing platforms were able to discriminate mismatches in the target sequences, theoretical detection limits to target DNA concentrations were calculated between 0.01 pM and 18.8 pM, improving resistance to non-specific interactions with DNA and proteins, fulfilling the requirements for a selective, sensitive, mobile, cost effective miniaturised DNA sensor.
Since four decades, rapid detection and monitoring in clinical and food diagnostics and in environmental and biodefense have paved the way for the elaboration of electrochemical biosensors. Thanks to their adaptability, ease of use in relatively complex samples, and their portability, electrochemical biosensors now are one of the mainstays of analy
This book covers the emerging topic of DNA nanotechnology and DNA supramolecular chemistry in its broader sense. By taking DNA out of its biological role, this biomolecule has become a very versatile building block in materials chemistry, supramolecular chemistry and bio-nanotechnology. Many novel structures have been realized in the past decade, which are now being used to create molecular machines, drug delivery systems, diagnosis platforms or potential electronic devices. The book combines many aspects of DNA nanotechnology, including formation of functional structures based on covalent and non-covalent systems, DNA origami, DNA based switches, DNA machines, and alternative structures and templates. This broad coverage is very appealing since it combines both the synthesis of modified DNA as well as designer concepts to successfully plan and make DNA nanostructures. Contributing authors have provided first a general introduction for the non-specialist reader, followed by a more in-depth analysis and presentation of their topic. In this way the book is attractive and useful for both the non-specialist who would like to have an overview of the topic, as well as the specialist reader who requires more information and inspiration to foster their own research.
Paper Based Sensors, Volume 89, the latest release in this comprehensive series that gathers the most important issues relating to the design and application of these cost-effective devices used in many industries, including health and environment diagnostics, safety and security, chemistry, optics, electrochemistry, nanoscience and nanotechnologies, presents the latest updates in the field. Chapters in this new release include Exploring paper as a substrate for electrochemical micro-devices, Paper-based sensors for application in biological compound detection, Printed paper-based (bio)sensors: design, fabrication and applications, Paper-based electrochemical sensing devices, Multifarious aspects of electrochemical paper-based (bio)sensors, Paper Based Biosensors for Clinical and Biomedical Applications, and more. Provides updates on the latest design in paper-based sensors using various nano and micromaterials Includes optical/electrical-based detection modes integrated within paper-based platforms Covers applications of paper-based platforms in diagnostics and other industries